Imaging system with a single color image sensor for simultaneous fluorescence and color video endoscopy

ABSTRACT

An endoscopic video system and method using a camera with a single color image sensor, for example a CCD color image sensor, for fluorescence and color imaging and for simultaneously displaying the images acquired in these imaging modes at video rates in real time is disclosed. The tissue under investigation is illuminated continuously with fluorescence excitation light and is further illuminated periodically using visible light outside of the fluorescence excitation wavelength range. The illumination sources may be conventional lamps using filters and shutters, or may include light-emitting diodes mounted at the distal tip of the endoscope.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/930,225, filed Jun. 28, 2013, now U.S. Pat. No. 9,143,746, which is acontinuation of U.S. application Ser. No. 11/964,330, filed Dec. 26,2007, now U.S. Pat. No. 8,498,695, which claims the benefit of U.S.Provisional Application No. 60/876,597, filed Dec. 22, 2006, and U.S.Provisional Application No. 60/908,373, filed Mar. 27, 2007, thedisclosures of all of which are incorporated herein by reference as iffully set forth herein.

BACKGROUND OF THE INVENTION

The invention is directed to methods and systems for simultaneousreal-time fluorescence and color video endoscopy at close to video framerates. The invention is also directed to high-efficiency illuminationsources and to methods and systems for controlling temporal and spectraloutput of these light sources.

Medical endoscopy is increasingly employing specialized optical imagingtechniques, such as fluorescence (i.e. autofluorescence andphotodynamic) endoscopy, narrow band imaging and other techniques, forimproved visualization and for the detection and diagnosis of diseases,Endoscopic imaging systems that provide specialized imaging modestypically also operate in a conventional color, or white-light,endoscopy mode. Embodiments of endoscopic imaging systems incorporatingboth a color and fluorescence imaging modes have been disclosed, forexample, in U.S. Pat. No. 6,462,770 B1, U.S. Pat. No. 6,821,245 B1, andU.S. Pat. No. 6,899,675 B2.

In conventional white-light endoscopy, hereinafter also referred to ascolor imaging mode, light in the visible spectral range is used toilluminate the tissue surface under observation. Light reflected by thetissue passes through a suitable lens system and is incident on an imagesensor built into or attached to the endoscope. The electrical signalsfrom the image sensor are processed into a full color video image whichcan be displayed on a video monitor or stored in a memory. Influorescence endoscopy, fluorescence excitation light excitesfluorophors in the tissue, which emit fluorescence light at an emissionwavelength which is typically greater than the excitation wavelength.Fluorescence light from the tissue passes through a suitable lens systemand is incident on the image sensor. The electrical signals from theimage sensor are processed into a fluorescence video image which can bedisplayed on a video monitor, either separately from or together withthe color video image, or stored in a memory.

The fluorescence excitation and emission wavelengths depend upon thetype of fluorophors being excited. In the case of exogenously appliedfluorophors, the band of excitation wavelengths may be located anywherein the range from the ultraviolet (UV) to the near infra-red (NIR) andthe emission wavelength band anywhere from the visible to the NIR. Forfluorophors endogenous to tissue, the band of excitation and emissionwavelengths are more limited (excitation from the UV to the green partof the visible spectrum, emission from the blue-green to the NIR).

In a conventional fluorescence/white-light endoscopic imaging system,the system can be switched between color and fluorescence modes eitherautomatically or with a hand- or foot-operated external switch. Both theillumination and imaging characteristics of the endoscopic imagingsystem may require adjustment when switching the operation of anendoscopic imaging system from one mode to the other. For example, gainadjustments and additional image processing (e.g., pixel binning, timeaveraging, etc.) may be required because the image signal in colorimaging mode tends to be substantially greater than the image signalfrom endogenous (tissue) fluorescence. Although switching betweenimaging modes with an automated device is not difficult, additional timemay be required to complete the endoscopic procedure because areas ofinterest are examined sequentially in each mode.

It would therefore be desirable to provide an endoscopic imaging systemcapable of acquiring and displaying images in both conventional color(“white-light”) and fluorescence imaging modes simultaneously. It wouldfurther be desirable to employ high-efficiency illumination sources thatcan be easily controlled over the spectral range of interest forendoscopy.

SUMMARY OF THE INVENTION

The invention disclosed herein describes an endoscopic video system andmethod using a single color image sensor for fluorescence and colorimaging and for simultaneously displaying the images acquired in theseimaging modes at video rates. The color imager may include a CCD colorimage sensor. The endoscopic video system has no moving parts.

According to one aspect of the invention, tissue is illuminatedcontinuously with fluorescence excitation light and is furtherilluminated periodically using visible light outside of the fluorescenceexcitation wavelength range. The method furthermore utilizes anexcitation light blocking filter which substantially blocks theexcitation light while allowing the blue, green and red components ofthe illumination light to pass to the color image sensor. In oneembodiment, the single color image sensor may be disposed in the tip ofthe endoscope, in which case the excitation light blocking filter ismounted in or on the tip of video endoscope.

With the method of the invention, fluorescence images are acquiredduring a time period when only the excitation light is supplied asillumination, while color images are acquired during a time period whenthe combination of both excitation light and visible light outside ofthe excitation wavelength range are supplied as illumination. The imagefields are read out from the single CCD color image sensor in aninterlaced fashion and processed to produce corresponding full-framefluorescence and white-light images. Real-time fluorescence andwhite-light images of the tissue are then produced by subtracting fromeach full-frame combined fluorescence and white-light image thecorresponding fluorescence image on a pixel-by pixel basis.

In one embodiment, the illumination light may be switched on for onecycle and switched off for two cycles, wherein a different image fieldof the combined tissue fluorescence and white-light image is read outduring each of the two cycles when the illumination light is switchedoff, and a different image field of the tissue fluorescence image areread out during each of the cycles when the illumination light isswitched on. A cycle may have a duration of 1/60 second. Four full framewhite-light images and two full frame fluorescence images may begenerated every six cycles.

The image data can be interpolated during cycles when no actual imagedata are available. For example, during a cycle where no full framewhite-light image is produced, an interpolated full frame white-lightimage may be computed from two adjacent full frame white-light images.Likewise, the fluorescence signals may be interpolated betweensequential fluorescence frames before being subtracted from thewhite-light image signals.

In yet another embodiment, pixel values of adjacent rows of the CCDcolor image sensor are added pixel-by-pixel to form summed row pixelvalues and the summed values are read out in an interlaced fashion.

In one embodiment, a high-resolution video image may be generated bycomputing a luma image of the combined full-frame fluorescence andwhite-light image signals and colorizing the luma image based on a ratioof red reflectance to fluorescence signals to produce a superimposedfluorescence/color image for display. Processing an image based on theluma data enhanced the attainable spatial resolution. A change in tissuepathology, as indicated by a change in the fluorescence signal from thattissue, can be represented as a change in color in the video image.

Further features and advantages of the present invention will beapparent from the following description of preferred embodiments andfrom the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

The following figures depict certain illustrative embodiments of theinvention in which like reference numerals refer to like elements. Thesedepicted embodiments are to be understood as illustrative of theinvention and not as limiting in any way.

FIG. 1 shows a schematic block diagram of an exemplary fluorescenceendoscopy video system with a single distal color image sensor;

FIG. 2 shows the camera of FIG. 1 with an excitation light blockingfilter;

FIG. 3 shows a schematic block diagram of a first exemplary embodimentof an illumination source according to the invention;

FIG. 4 shows a schematic block diagram of a second exemplary embodimentof an illumination source according to the invention;

FIGS. 5A-5C show a filter arrangement on a CMGY image sensor (FIG. 5A)and interlaced readout (FIGS. 5B-5C) with summing on chip;

FIGS. 6A-6C show a filter arrangement on a CMGY image sensor (FIG. 6A)and interlaced readout (FIGS. 6B-6C) without summing on chip;

FIG. 7 shows a timing diagram for excitation light and imaging lightexposure;

FIG. 8 shows a timing diagram for reading from the color sensorfluorescence and color image information;

FIG. 9 shows schematically block diagram of a process according to theinvention for extracting fluorescence and color images; and

FIGS. 10A and 10B illustrate an LED assembly configured as amulti-wavelength illumination source for endoscopy.

DETAILED DESCRIPTION

In conventional white-light (color imaging) endoscopy, broadband visiblelight is used to illuminate the tissue under observation. Historically,endoscopes used for white light endoscopy have incorporated fiberopticlight guides to transmit light from lamps to provide this type ofillumination. In fluorescence endoscopy, fluorophors in the tissue areexcited by illumination with a shorter wavelength light and theresulting fluorescence emission is detected at Stokes-shifted longerwavelengths. The fluorophors may be either endogenous to the tissue(i.e., naturally present) or exogenous (e.g., dyes applied to enhancecontrast for diagnostic or other imaging purposes). Since thefluorescence process tends to be rather inefficient, the intensity ofthe shorter wavelength excitation light is typically several orders ofmagnitude greater than the intensity of the resulting fluorescenceemission. As such, both direct visualization and imaging of emissionsfrom fluorophors requires the use of a barrier filter that blockstransmission of the reflected shorter wavelength excitation light andprevents the excitation light from overwhelming the eye or image sensorused to observe/detect the emitted fluorescence. A certain minimum levelof excitation light intensity is also required to provide the desiredquality of (optical or electronic) image signal. The desired amount ofexcitation light will depend on the type and concentration offluorophors to be excited, distance to the tissue and size of the areabeing visualized imaged, the sensitivity of the eye/image sensor andsimilar related factors. As a result, particularly in the case ofnatural (i.e., endogenous) tissue fluorescence, endoscopy imagingsystems operating in fluorescence mode typically employ powerful arclamps or lasers to excite fluorophors as well as highly sensitivecameras to image fluorescence emissions from these fluorophors.

FIG. 1 is a block diagram of a fluorescence endoscopy video system 1 inaccordance with one embodiment of the present invention. The systemincludes a multi-mode light source 12 that generates light for obtainingcolor and fluorescence images. The use of the light source for obtainingdifferent kinds of images will be described in further detail below.Light from the light source 12 is supplied to an illumination guide 16of an endoscope 10, which then illuminates a tissue sample 200 that isto be imaged.

As also shown in FIG. 1, the system also includes a camera 100, forexample, a solid-state camera based on a CCD or CMOS sensor chip, whichin the exemplary embodiment is located at the distal or insertion end ofthe endoscope 60. Alternatively, although not illustrated, the camera100 may also be positioned at another location, such as the proximal endof the endoscope 60. In the depicted embodiment, the light from thetissue is directly captured by the camera 100, and the operation of thesystem is similar to video endoscopes currently on the market (such asthe Olympus CF-240L).

A processor/controller 14 controls the camera 100 and the light source12, which will be described in more detail below, and produces videosignals that are displayed on a video monitor 18. Theprocessor/controller 14 communicates with the camera 100 by wire orother signal communication devices that are routed within the endoscope,such as optical fiber. Alternatively, communication between theprocessor/controller 14 and the camera 100 can be conducted over awireless link. Clinically relevant information about the health of thetissue under observation may be contained in the intensity of thefluorescence emission within a specific wavelength range.

For autofluorescence endoscopy (endoscopy using endogenous fluorophors),such information is contained in the green wavelength range of theemitted fluorescence. It has been observed that green florescence isincreasingly suppressed as the tissue becomes increasingly diseased.However, the red fluorescence signal does not vary with the diseasestate of the tissue and can hence be used to distinguish betweenintensity variation in the green fluorescence emission due to thedisease state of the tissue and intensity variations due to imagingartifacts, such as shadows or geometry effects (e.g., imaging distance).A single multicolor image can be formed in which the color is indicativeof the health of the examined tissue by combining the image informationfrom a wavelength range that varies with the disease state (greenfluorescence) with the image information from a wavelength range thatdoes not vary with the disease state (red fluorescence) of the tissue.

FIG. 2 shows schematically an exemplary embodiment of camera 100 withcolor image sensor 22 and light collimating optics 26. Positionedbetween the tissue 200 and color image sensor 22 is an excitation lightblocking filter 24 which blocks reflected excitation light from reachingimage sensor 22, while allowing imaging light and fluorescence light topass. The advantage of this configuration is that all imaging isperformed and controlled by the same imaging optics 113. In analternative embodiment, the excitation light blocking filter 24 may beplaced distal of the light collimating optics 26, and in someembodiments may be disposed on the outside of the distal tip of theendoscope, for example, when converting a white-light imaging endoscopeinto an imaging/fluorescence endoscope. An externally mounted excitationlight blocking filter is described in, for example, commonly assignedU.S. application Ser. No. 11/412,715.

The white light/fluorescence video endoscopy system of the inventionoperates by illuminating the sample with either excitation light aloneor with a combination of excitation light and illumination light in awavelength range or in wavelength ranges outside the spectral range ofthe excitation spectrum. The light source for excitation light andillumination light can be, for example, an arc lamp, a solid state lightemitter such as one or more diode lasers or light emitting diodes, orany other light source emitting light in a suitable wavelength range.The light source can be a single light source, wherein a portion of thelight is filtered out to provide excitation light, and another portionof the light is filtered out to provide illumination light.Alternatively, different light sources can be provided for excitationlight and illumination light, respectively. The illumination light istimed, either by using an external shutter 37 or, if light sources witha rapid response are used, by turning the light sources on and off.

FIG. 3 shows in more detail a first embodiment of a multi-mode lightsource 30 for simultaneously illuminating a tissue sample 200 withcontinuous fluorescence excitation light and switched illuminationlight. Light source 30 includes a first light source 31, for example, anarc lamp, and a collimating lens 33 for producing a high intensity,preferably collimated spectral output S_(source) which includes anexcitation wavelength range. A bandpass filter 34 filters out spectralcomponents outside the excitation wavelength range S_(excitation) andallows only spectral components within the excitation wavelength rangeS_(excitation) to pass. Light source 30 further includes a second lightsource 32, for example, a halogen lamp, for producing a preferablycollimated spectral output S_(illumination) with a high intensity in animaging wavelength range covering, for example, the visible spectralrange. Light source 32 may be switched with timing signals produced byprocessor/controller 14, for example, by placing a mechanical orelectronic shutter 37 between second light source 32 and dichroic mirror35 or by controlling the electric current supplied to light source 32.The combined collimated excitation/imaging light is focused by lens 36onto the input face 37 of an optical fiber illumination guide 16 with asuitable numerical aperture (NA).

FIG. 4 shows a second embodiment of a multi-mode light source 40 forsimultaneously illuminating a tissue sample 200 with continuousfluorescence excitation light and switched illumination light. Lightsource 40 includes an excitation/illumination light source 31, forexample, an arc lamp, and a collimating lens 33 for producing a highintensity, preferably collimated spectral output S_(source) whichincludes an excitation wavelength range S_(excitation). A dichroicmirror 41 reflects the spectral illumination component S_(illumination)and passes the excitation wavelength range S_(excitation) which may beadditionally narrow-band filtered by bandpass filter 34. The lightcomponent reflected off a first dichroic mirror 41 is then reflected bymirror 42, passes through a shutter 45 (mechanical or electronic) and isthen further reflected by mirror 43 and reflected at second dichroicmirror 44 to become collinear with the excitation light passing throughfilter 34. As before, the combined collimated excitation/imaging lightis focused by lens 36 onto the input face 37 of an optical fiberillumination guide 16 with a suitable numerical aperture (NA). Thisembodiment takes advantage of the fact that a suitable arc lamp can emitover a wavelength range which covers both the excitation light spectrumand the illumination light spectrum. The shutter 45 may be switched bytiming signals produced by processor/controller 14.

Suitable filters, for example, a low-pass filter to block excitationlight and/or a high-pass filter to block unswitched illumination light,may be placed along the optical paths.

In operation, when the switched light source 32 is off (or the shutter45 is closed), only excitation light illuminates the tissue 200, forexample, through the endoscope illumination guide 16. The reflectedexcitation light is blocked from reaching the color image sensor by theexcitation light blocking filter 24, while tissue fluorescence lightpasses through the excitation light blocking filter 24 and reaches thecolor image sensor 22 for fluorescence light detection.

When the illumination light source 32 is switched on (or the shutter 45is open), the combined light from the illumination light source 32 andthe excitation light source 31 is coupled into the endoscope light guide14 and illuminates the tissue 200. The reflected excitation light (andany residual light from the switched light source at that wavelength) isblocked as before by the excitation light blocking filter 24, while thecombination of both tissue fluorescence and reflected illumination light(“white light”) is imaged by the color image sensor 22.

FIGS. 5A-5C show an exemplary arrangement of spectral filter elementsdisposed on the pixels of a CMGY image sensor (FIG. 5A) and aninterlaced readout (FIGS. 5B-5C) with on-chip summing of pixels fromadjacent rows. The first half-frame in the embodiment depicted in FIGS.5A, 5B, 5C is here composed of the sum of lines 1 and 2; 3 and 4; 5 and6; and so on, whereas the second half-frame is composed of the sum oflines 2 and 3; 4 and 5; and so on. FIG. 6A shows the same filterarrangement as in FIG. 5A, but with a different interlaced readout(FIGS. 6B-6C) without on-chip summing. The first half-frame in theembodiment depicted in FIGS. 6A, 6B, 6C is composed of lines 1; 3; 5;and so on, whereas the second half-frame is composed of the lines 2; 4;and so on.

Most video endoscopes and endoscopic video cameras currently use CODimage sensors with CMGY color filters since these tend to provide thehighest quality color images.

FIG. 7 shows a timing diagram according to the invention for operatingthe exemplary endoscope system. As can be seen from curve (a) in thediagram, the excitation light source 31 is turned on at time T₀,irradiating the tissue continuously with fluorescence excitation light.Conversely, as depicted by curve (b), the illumination light source 32is periodically switched on and off (or shutter 37 or 45 is opened andclosed) with a duty factor of 33%, i.e. the illumination light source isturned on at times T₁, T₄, T₇, . . . (i.e., at times T_(1+3*n) with n=0,1, 2 . . . ) for one field period and turned off again at times T₂, T₅,T₈, . . . , T_(2+3*n) for two field periods, which include times T₃, T₆,T₉, . . . , T_(3+3*n). In the depicted example, a field period has aduration of 1/60 s=16.7 ms.

As mentioned above, the exemplary image sensor is read out in aninterlaced fashion, so that even lines and odd lines are readalternatingly, with or without summation on the chip. An image with fullvertical resolution is then generated in the video processor/controller14 by combining two sequential interlaced fields to form a full videoframe for the fluorescence image and for the combinedfluorescence/white-light image.

FIG. 8 describes in more detail the temporal illumination and readoutpattern of the interlaced CCD image as a function of time.

Before the image acquisition begins in the depicted example at time T₁,the COD is illuminated only with fluorescence excitation light. Theeven-fields acquired in the time interval preceding T₁ containfluorescence-only data which are read out at T₁. At the same time, theillumination light is turned on, so that the COD is now illuminated withfluorescence excitation light and illumination light between the timesT₁ and T₂.

The illumination light is turned off at time T₂, in the present exampleafter 16.7 ms, and the image data representing “color-plus-fluorescence”are read out for the odd field at T₂ and for the even field at T₃. TheCOD is illuminated from T₂ until T₄ with fluorescence light only andacquires a new fluorescence signal. It should be noted that thefluorescence signal is acquired during two field periods, whereas theadded illumination light is acquired only during one field period, whichprovides an improved signal over other methods, where the fluorescencesignal and the illumination signal are acquired with the same dutycycle.

The image signals from the color image sensor acquired alternatinglyduring “fluorescence-only” and “color-plus-fluorescence” measurementsare supplied to processor/controller 14 which stores and processes theimage signals to form the desired images for display. Theprocessor/controller 14 may be, for example, a processor selected fromthe Texas Instruments C64XX family of image processors. The processingof a specific field depends on whether the field is to be used togenerate a fluorescence image or a color (white tight) image. Theprocessor/controller 14 may also synchronize the operation of theswitched illumination light source with the image acquisition, asdescribed above.

This exposure and read-out scheme described above generates from thecombination of odd and even fields a full frame of fluorescence imageinformation every six field time periods. In the depicted example, eachfield time period is 16.7 ms. In other words, the full framefluorescence image is completely updated every tenth of a second. Duringthe same six (6) field periods, four fields (two even fields and two oddfields) of color image information are generated and these even- andodd-line fields are suitably combined and processed to generate four (4)full vertical resolution color video frames during the same six (6)field periods. As seen in column 6 of FIG. 8, the display signal writteninto buffer memory still includes the fluorescence signal component,which is then subtracted to yield the color image signal. Thetransformation of image data from the CMGY image space to the RGB imagespace for display is conventional and will not be described further.

Because during six (6) field periods the image data contain only 2 (two)fields of color information, rather than three (3) video frames, theimage data may advantageously be interpolated between sequential datapoints. In this way, the image quality can be improved by providing asmooth transition between frames, so that the final color video image isperceived by the human eye as being substantially similar to the fieldupdate rate in a normal video signal.

Once the image signals in Column 6 of FIG. 8 are transferred to theimage processor 14, the color (white-light) images and the fluorescenceimages are separated on a frame-by-frame basis. The color information isextracted from these frames (i.e. the contribution from the fluorescencesignal is removed) by subtracting pixel-by-pixel a fluorescence signalvalue from each “color-plus-fluorescence” frame. Advantageously, thesubtracted fluorescence signal values are interpolated from thepreceding stored “fluorescence-only” frame and the “fluorescence-only”frame following the “color-plus-fluorescence” frame being processed.This causes at most a delay of two fields, in the present example of66.7 ms, between image acquisition and display.

After the fluorescence contribution is subtracted, the color balance ofthe remaining image signals may still need to be corrected for properwhite balance. This correction may be performed using conventional imageprocessing and color-space transformation methods by using acompensation matrix or similar processing techniques, which convert theimage signal from one color space to another. The processing offluorescence image fields is somewhat less complex because thefluorescence image data do not include image data from other sources.Accordingly, fluorescence image data produced in multiple,non-overlapping spectral ranges may be processed and displayed as a realcolor or false color image (for example, green fluorescence fromfluorescein) may be displayed as green and IR fluorescence from ICG maybe displayed as red, etc., in the same fashion as white light colorimages are processed and displayed on a video monitor. Using this typeof fluorescence imaging display for autofluorescence or endogenoustissue fluorescence imaging, areas of tissue in which the greenfluorescence is suppressed due to abnormal pathology will appear redsince the red fluorescence is proportionally less suppressed.

The processor/controller circuit 14 can carry out inter-imagecomputation for superimposing a fluorescence image and a white-lightlight image on video monitor 18. An operator can therefore view thefluorescence image and the white-light light image simultaneously,without introducing a perceptible time delay between them. Consequently,for example, the location of a lesion can be readily viewed with highprecision, which is very useful for diagnosis.

FIG. 9 illustrates schematically a process flow which may be performed,for example, by processor/controller 14, to extract fluorescence imagesand reflectance images, and to correct image intensity and colorinformation for improving spatial resolution and for simultaneouslydisplaying fluorescence/reflectance images.

The depicted process assumes that the excitation light, labeled (A) inFIG. 9, is emitted in the blue/UV spectral range, for example, forexciting fluorescence in fluorescein, which is detected in the greenspectral range. However, other fluorescent dyes such as ICG which hasexcitation/fluorescence wavelengths in the red/IR spectral range canalso be used, and the present invention is not limited to particularfluorescent materials or wavelengths. Illumination light is emitted atwavelengths outside the excitation light wavelengths and is showntogether with the excitation light in FIG. 9 as (B).

When the tissue is illuminated with fluorescence light only, e.g.,during the time interval between T₀ and T₁ (FIG. 7), a fluorescencespectrum (C) is detected by color sensor 22. When the tissue isilluminated with fluorescence light+illumination light, e.g., in thetime interval between T₁ and T₂ (FIG. 7), a fluorescence+color imagespectrum (D) is detected by color sensor 22. The fluorescence spectrum(C) is then subtracted from the fluorescence+color image spectrum (ID)to produce the spectral response of the color image (E). This colorimage can then be displayed at 92.

Advantageously, the “luma” component of the fluorescence+color image isextracted, shown as (F). Luma refers to the brightness in an image,i.e., the not-gamma-corrected “black and white” or achromatic portion ofthe image. Stated differently, luma represents the achromatic imagewithout any color, while the chroma components represent the colorinformation. The luma component can be used for extracting more accuratespatial information from the image data.

In one embodiment, the red reflectance signal (G) is extracted from thecolor image frames. A ratio of fluorescence to red reflectance forspatially corresponding pixels in the fluorescence and color videoframes is calculated, at 94, on a pixel-by-pixel basis, and the value ofthat ratio is used to determine the color (chroma) of the display pixelat that same location, at 94. The color of a display pixel is assignedsuch that ratio values that indicate suppressed green fluorescence andabnormal pathology are rendered in a contrasting color to pixels inwhich the ratio values are characteristic of normal green fluorescencevalues indicating normal tissue pathology. Although the color (chroma)of the display pixels is based upon a ratio of fluorescence toreflectance signal for that pixel, the brightness (luma) of each displaypixel may simply be taken as the brightness (luma) of each color videoframe pixel. Because the color, or white-light, video fields are updatedat near video rates (i.e. 4 times in a 6 field period, see FIGS. 7 and8), the resulting fluorescence/reflectance image brightness defining theluma will also be updated at that rate. Conversely, the chroma portionof the fluorescence/reflectance image will be updated somewhat moregradually (due to the less frequent field update rate of thefluorescence image signals). However, the human eye is less sensitive tochanges in color than to changes in brightness, so that the slowerfluorescence field update rate will be less objectionable in the imagedisplay and can still be regarded as a real-time image. The luma image(F) can then be colored according to the chroma information derived fromthe red reflectance (G).

Normalizing a fluorescence image by a red light image is advantageous,because the color of mucosa inside a human body is dominated byhemoglobin which is a pigment and predominantly absorbs light withwavelengths shorter than 600 nm. The reference image used fornormalization should therefore represent reflected wavelengths of 600 nmor longer. The normalized fluorescence image can then be used as anaccurate representation of the intensity of actual fluorescence or thedegree of accumulation of an antibody labeled, for example, byindocyanine green (ICG). Normalization of a fluorescence image is notlimited to normalization relative to a red light image. Alternatively,an image depicted by infrared fluorescence components may be used forthe normalization.

It should be mentioned that for removing excitation light, theexcitation light blocking filter 24 in FIG. 2 may be replaced by adichroic mirror which reflects the spectral components of the excitationlight.

Recent developments in solid state lighting technology have given riseto the use of solid state devices, such as light-emitting diodes (LEDs)and lasers, as sources of endoscopic illumination which may eventuallyreplace the lamps 31 and 32 in the multimode light source 12. Since LEDsare very compact, inexpensive, reliable, and have a long lifetime (onthe order of 10,000 hours or longer, depending on the drive current),incorporation of this illumination technique in endoscopic medicalequipment will lead to lower cost endoscopic light sources and hencealso to less expensive endoscopes.

Solid state illumination sources, in particular LEDs, with emissionwavelengths ranging from the deep UV to the infrared spectral range,have recently become available. These LEDs have several advantages whichmakes them particularly suitable for endoscopy: they can be manufacturedto have a narrow, controllable spectral emission range which may betuned to the fluorescence excitation spectra of the fluorophors; theyare very efficiently in converting electric input power to opticaloutput power; they can be rapidly switched on and off; and their poweroutput can be adjusted by varying the electric current through thedevice which facilitates control and timing of the spectral output of anLED-based illumination source.

Due to their small die size, LEDs may be disposed at or incorporated inthe distal tip of an endoscope. For example, as shown schematically inFIGS. 10A and 10B, several LEDs mounted on a common carrier can provideboth narrow-band shorter wavelength excitation light for fluorescenceendoscopy and broader visible illumination light for white-lightendoscopy. FIG. 10A is a schematic top view of an illumination assembly110 with an excitation, e.g. UV LED 112 die for providing excitationlight, which is surrounded by blue (CWL 470 nm), green (CWL 525 nm),(CWL 590 nm) amber and red (CWL 630 nm) LED dies 114, 115, 116, 117 thatprovide illumination light. The indicated wavelengths are exemplary onlyand not intended to limit the scope of the invention. Also indicated arebonding pads 122 to electrically connect the LEDs to external wires (notshown). In general, more than the two indicated bonding pads may beprovided. Each of the LEDs may be controlled individually.

In another embodiment not shown in the drawings, a so-called “white” LEDwhich generates illumination light covering the visible spectral rangecan be employed instead of separate blue, green, red, and amber LEDs.“White” LEDs convert blue or UV radiation emitted by the blue- orUV-emitting LED die to visible light by downconversion of the blue- orUV-emission with a suitable phosphor. Both types of LEDs have recentlybecome commercially available. Advantageously, the LEDs can be lensedfor efficient directional illumination of the target tissue. Theexcitation LED may emit light in any spectral range suitable forexciting fluorescence in a dye, such as in the blue for fluorescein andin the near IR for ICG.

It will be understood that light emitted by the illumination LEDs shouldnot contain spectral components in a wavelength range where dyefluorescence is excited. To eliminate emission at excitation lightwavelengths from reaching the tissue under examination, suitable cutoffor passband, for example notch filters, may be placed in the opticalpath of the separate color LEDs or the “white-light” LEDs ofillumination assembly 110.

Although LEDs convert electric energy to optical energy veryefficiently, they still generate a substantial amount of heat which maycause discomfort for the patient. These LEDs may therefore have to becooled. As shown more clearly in FIG. 10B, the LEDs may be mounted on aheat sink 118 with a coolant inlet/outlet which can be connected to anexternal chiller. In general, devices for cooling the LEDs may includethermoelectric coolers, liquid-cooled heat exchangers, expansioncoolers, microchannel coolers, thermo-siphon heat pipes, and the like.

The excitation light blocking filter 24 for the excitation light placedin front of the sensor may be designed to prevent transmission of blueor UV light produced by the white-light LED. Alternatively or inaddition, the LED itself may be covered with a filter absorbing the blueor UV light from the LED dies.

A temperature sensor may be incorporated into the heat sink 118, ormounted in close vicinity to the LED array, for the purposes of

1. monitoring and adjusting the heat sink temperature, and

2. providing a safety mechanism by which a signal can be generated toreduce or interrupt the electrical power to the LEDs in the event of afailure in the heat sink cooling system.

While the invention has been disclosed in connection with the preferredembodiments shown and described in detail, various modifications andimprovements thereon will become readily apparent to those skilled inthe art. For example, although not illustrated in the drawings, theillumination sources, such as the arc lamp or halogen lamp, may bereplaced with LEDs or lasers. Accordingly, the spirit and scope of thepresent invention is to be limited only by the following claims.

What is claimed is:
 1. A method for visualizing a tissue of a subject,the method comprising: illuminating the tissue with a white light and anexcitation light that excites fluorophors in the tissue, wherein thefluorophors emit fluorescence light to create a fluorescence image;continuously acquiring fluorescence and white light reflectance imagesof the tissue; and displaying images of the tissue generated from thecontinuously acquired fluorescence and white light reflectance images atvideo frame rates on a display device, wherein generating the displayedimages comprises: receiving a fluorescence image of the tissue and awhite light reflectance image of the tissue that is formed fromreflectance of the illuminated white light, wherein the fluorescence andthe white light reflectance images have spatially corresponding pixels;calculating, for each of the spatially corresponding pixels in thefluorescence and reflectance images on a pixel-by-pixel basis, a ratiobetween a fluorescence signal for each pixel in the fluorescence imageand an extracted color reflectance signal from the white lightreflectance image for each pixel in the reflectance image; andgenerating an image of the tissue, wherein each pixel in the generatedimage has a brightness based on brightness of its corresponding pixel inthe white light reflectance image, and wherein each pixel in thegenerated image is assigned a color based on the calculated ratio forits corresponding pixel in the fluorescence and reflectance images,wherein the assigned colors in the generated image comprise a firstcolor indicating a first tissue characteristic and a second colorindicating a second tissue characteristic.
 2. The method of claim 1,wherein the first and second colors are contrasting.
 3. The method ofclaim 1, wherein the first tissue characteristic is abnormal tissuepathology and the second tissue characteristic is normal tissuepathology.
 4. The method of claim 1, wherein the extracted colorreflectance signal for each pixel is a red reflectance signal.
 5. Themethod of claim 1, wherein the calculated ratio is the ratio of thefluorescence signal to the extracted color reflectance signal.
 6. Themethod of claim 1, wherein the calculated ratio is the ratio of theextracted color reflectance signal to the fluorescence signal.
 7. Themethod of claim 1, wherein the reflectance image and the fluorescenceimage have been produced from a combined reflectance and fluorescenceimage.
 8. The method of claim 1, wherein a sensor used to receive thereflectance image is also used to receive the fluorescence image.
 9. Themethod of claim 1, wherein receiving the reflectance image and thefluorescence image comprises receiving a combined reflectance andfluorescence signal.
 10. The method of claim 1, wherein the receiving isperformed using an endoscope.
 11. The method of claim 1, furthercomprising displaying images of the tissue generated from thecontinuously acquired fluorescence and white light reflectance images atvideo frame rates in real-time.
 12. A system for visualizing a tissue ofa subject, the system comprising: a light source that providesfluorescence excitation light to excite fluorophors in the tissue,wherein the fluorophors emit fluorescence light to create a fluorescenceimage, and white light reflectance illumination light; a camera thatcontinuously acquires white light reflectance images of the tissue thatare formed from reflectance of white light from the white lightreflectance illumination source and fluorescence images of the tissue,wherein the reflectance and fluorescence images have spatiallycorresponding pixels; and a processor in communication with the camerathat continuously receives the fluorescence and reflectance images;calculates, for each of the spatially corresponding pixels in thefluorescence and reflectance images on a pixel-by-pixel basis, a ratiobetween a fluorescence signal for each pixel in the fluorescence imageand an extracted color reflectance signal based on the white lightreflectance image for each pixel in the reflectance image; and generatesimages of the tissue at video frame rates, wherein each pixel in thegenerated images has a brightness based on brightness of itscorresponding pixel in the white light reflectance image, and whereineach pixel of the generated images is assigned a color based on thecalculated ratio for its corresponding pixel in the fluorescence andreflectance images, wherein the assigned colors in the generated imagescomprise a first color indicating a first tissue characteristic and asecond color indicating a second tissue characteristic.
 13. The systemof claim 12, wherein the first and second colors are contrasting. 14.The system of claim 12, wherein the first tissue characteristic isabnormal tissue pathology and the second tissue characteristic is normaltissue pathology.
 15. The system of claim 12, further comprising adisplay device that simultaneously displays the generated images of thetissue.
 16. The system of claim 12, wherein the extracted colorreflectance signal is a red reflectance signal.
 17. The system of claim12, wherein the calculated ratio is the ratio of the fluorescence signalto the extracted color reflectance signal.
 18. The system of claim 12,wherein the calculated ratio is the ratio of the extracted colorreflectance signal to the fluorescence signal.
 19. The system of claim12, wherein the camera comprises a sensor that is used to acquire thereflectance images and is also used to acquire the fluorescence images.20. The system of claim 12, wherein the camera is located at aninsertion end of an endoscope.
 21. The system of claim 12, wherein thecamera is located at a proximal end of an endoscope.
 22. The system ofclaim 19, wherein the sensor comprises a CMOS sensor chip.
 23. Thesystem of claim 12, wherein the light source comprises a light-emittingdiode that is switched on and off.
 24. A method for visualizing a tissueof a subject, the method comprising: administering a fluorescent dye tothe subject; illuminating the tissue with a white light and exciting thefluorescent dye in the tissue to excite fluorophors in the tissue,wherein the fluorophors emit fluorescence light to create a fluorescenceimage; continuously acquiring fluorescence and white light reflectanceimages of the tissue; and displaying images of the tissue generated fromthe continuously acquired fluorescence and white light reflectanceimages at video frame rates on a display device, wherein generating thedisplayed images comprises: receiving a fluorescence image of the tissueand a reflectance image of the tissue that is formed from reflectance ofthe illuminated white light, wherein the fluorescence and reflectanceimages have spatially corresponding pixels; calculating, for each of thespatially corresponding pixels in the fluorescence and reflectanceimages on a pixel-by-pixel basis, a ratio between a fluorescence signalfor each pixel in the fluorescence image and an extracted colorreflectance signal for each pixel in the reflectance image; andgenerating an image of the tissue, wherein each pixel in the generatedimage has a brightness based on brightness of its corresponding pixel inthe white light reflectance image, and wherein each pixel in thegenerated image is assigned a color based on the calculated ratio forits corresponding pixel in the fluorescence and reflectance images,wherein the assigned colors in the generated image comprise a firstcolor indicating a first tissue characteristic and a second colorindicating a second tissue characteristic.
 25. The method of claim 24,wherein the first and second colors are contrasting.
 26. The method ofclaim 24, wherein the first tissue characteristic is abnormal tissuepathology and the second tissue characteristic is normal tissuepathology.
 27. The method of claim 24, wherein the extracted colorreflectance signal for each pixel is a red reflectance signal.
 28. Themethod of claim 24, wherein the calculated ratio is the ratio of thefluorescence signal to the extracted color reflectance signal.
 29. Themethod of claim 24, wherein the calculated ratio is the ratio of theextracted color reflectance signal to the fluorescence signal.
 30. Themethod of claim 24, wherein the reflectance image and the fluorescenceimage have been produced from a combined reflectance and fluorescenceimage.
 31. The method of claim 24, further comprising acquiring thereflectance image and the fluorescence image.
 32. The method of claim31, wherein a sensor used to acquire the reflectance image is also usedto acquire the fluorescence image.
 33. The method of claim 31, whereinacquiring the reflectance image and the fluorescence image comprisesacquiring a combined reflectance and fluorescence signal.
 34. The methodof claim 31, wherein the acquisition is performed using an endoscope.35. The method of claim 24, further comprising generating one or morefluorescence images by interpolation.
 36. The method of claim 24,further comprising generating one or more reflectance images byinterpolation.
 37. The method of claim 24, wherein the dye comprisesindocyanine green (ICG).
 38. The method of claim 24, wherein the dye isindocyanine green (ICG).
 39. The method of claim 1, further comprisingadministering a fluorescent dye to the subject comprising indocyaninegreen (ICG).
 40. The method of claim 39, wherein the dye is indocyaninegreen (ICG).